RU2434867C2 - Benzoquinolizine-2-carboxylic acid derivatives - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to novel benzoquinolizine-2-carboxylic acid derivatives of formula I and pharmaceutically acceptable solvates thereof in which: R denotes CH₃ or CH(CH₃)₂; and R₁ denotes C₁-C₆ alkyl or phenyl, substituted with one or more substitutes selected from C₁-C₆ alkyl or nitro if necessary. The invention also relates to a method of producing a compound of formula

I, a pharmaceutical composition based on the compound of formula I and use of the compound of formula I.

EFFECT: novel benzoquinolizine-2-carboxylic acid derivatives are obtained, having higher solubility in water in a wide pH range, which can be used to treat bacterial gram-positive, gram-negative and anaerobic infections.

18 cl, 2 tbl, 8 ex

Description

Field of Invention

The present invention relates to new prodrugs of optically pure benzoquinolizin-2-carboxylic acid and to pharmaceutical compositions that include these prodrugs. The compounds and compositions of the invention can be used to treat infections caused by gram-positive, gram-negative and anaerobic bacteria, especially infections caused by a resistant gram-positive and gram-negative organism, mycobacterial infections, and infections caused by nosocomial pathogens.

State of the art

Prodrugs are therapeutic agents that are inactive per se, but in vivo they transform into a therapeutically active parent molecule. Prodrugs have optimal physicochemical, pharmacokinetic and pharmacodynamic properties. They may be designed to overcome pharmaceutical, pharmacokinetic or pharmacodynamic barriers such as insufficient oral absorption, insufficient solubility, lack of chemical stability, unacceptable taste or smell, irritation or pain, inadequate passage through the blood-brain barrier, toxicity, and pronounced presystemic metabolism.

For the convenience of patients, most drugs are administered orally. There are significant problems that prevent the delivery of a drug from the oral route, which often means that not all of the administered compound reaches its intended site of action. The extent to which the compound can overcome the obstacles to the oral use of the drug and achieve a wide range of blood circulation is quantified by the term oral bioavailability.

Chiral fluoroquinolone S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolizin- 2-carboxylic acid, also known as S - (-) - nadifloxacin, is described in Japanese patents JP 63,192,753A and JP 05,339,238A.

S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2- carboxylic acid has the potential to be used as a commercial antibacterial agent due to its antibacterial activity profile. However, the sodium salt of S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolizine -2-carboxylic acid tends to cause phlebitis in rats when administered intravenously. Also S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2 β-carboxylic acid has a solubility in water of 0.8-2.0 mg / ml in the pH range of 8.0-9.5 at 28 ° C, thus creating problems in terms of the need to formulate the drug in tablet or capsule form or in the creation of formulations for artificial nutrition and parenteral injection. Thus, a desired agent that can overcome the above problems can help develop a dosage form suitable for systemic use in humans and animals.

In this direction, the authors showed that the salt S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2-carboxylic acid and L-arginine is a broad-spectrum antibiotic that has a lower tendency to induce phlebitis in rats when administered by the intravenous route, as disclosed in PCT application WO00 / 68229. The most stable salt form is the tetrahydrate of salt S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i , j] quinolysin-2-carboxylic acid and L-arginine, is disclosed in PCT application WO05 / 023805A1 (and in the corresponding US patent US 7 164 023). It is useful to obtain stable pharmaceutical dosage forms, including an aqueous solution. The injection form is particularly suitable for long-term intravenous therapy for the treatment of diseases caused by bacterial infections, due to its favorable solubility in water at pH 9.5, its ideal suitability because it does not cause venous inflammation upon repeated intravenous administration, and security profile.

However, it was found that the oral bioavailability of S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i , j] quinolysin-2-carboxylic acid and tetrahydrate of the salt S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2-carboxylic acid and L-arginine dimethylbenzoic acid is insufficient when used in animals. Poor oral bioavailability of S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolizine -2-carboxylic acid is probably the result of inadequate solubility, and therefore inadequate absorption in vivo . At the same time, the insufficient oral bioavailability of the salt of L-arginine and S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2-carboxylic acid is probably a consequence of the tendency of the compound to precipitate at acidic pH during transit through the gastric region, where pH is ~ 1-2.

Thus, to improve the oral bioavailability of S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i , j] quinolysin-2-carboxylic acid, a prodrug approach has been provided. Prodrugs S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2 β-carboxylic acid are disclosed in US Pat. No. 6,750,224 to the applicant. The '224 patent discloses two types of prodrugs — prodrugs in the 2-carboxyl group and in the 4-hydroxyl group of the piperidine side chain. Prodrugs disclosed in 4-hydroxypiperidine include amino acid prodrugs. These prodrugs have been studied to develop an oral pharmaceutical composition. Prodrugs S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2 β-carboxylic acid disclosed therein, or did not have optimal characteristics, such as adequate solubility in a wide pH range, which is likely to be present in the digestive tract, in order to demonstrate the real effect of a prodrug, or prodrugs were not able to improve oral bioavailability S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2- carboxylic sour compared with the tetrahydrate salts of S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2-carboxylic acid and L-arginine. Among the amino acid prodrugs, prodrugs of L-alanine and L-valine have also been disclosed. Both prodrugs had limited water solubility (<10 mg / ml). It is generally assumed that inadequate solubility in water is an important factor limiting oral bioavailability.

An object of the present invention is to provide a compound with improved water solubility over a wide pH range, which is likely to occur in the digestive tract, and with increased oral bioavailability.

SUMMARY OF THE INVENTION

According to the present invention, prodrug salts of S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2-carboxylic acid of Formula I

and their pharmaceutically acceptable solvates, polymorphs or hydrates,

wherein

R is CH ₃ or CH (CH ₃ ) ₂ and

R ₁ is C ₁ -C ₆ alkyl or phenyl, optionally substituted with one or more substituents selected from C ₁ -C ₆ alkyl, halogen, nitro, hydroxy or C ₁ -C ₆ alkoxy.

The present invention also relates to a method for producing a compound of formula I, comprising the steps of combining an amine protected L-alanine or L-valine with S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1- il) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2-carboxylic acid to obtain a compound of formula II, removing the protective group from the compound of formula II and isolating the compound of the invention of formula I or its pharmaceutically acceptable salt.

Additionally, this invention relates to a method or methods for producing compounds of the invention of formula I and their polymorphs and hydrates.

The invention also relates to pharmaceutical compositions containing the compounds of the invention, and to a method for treating or preventing bacterial infections using the compounds and compositions of the invention.

Details of one or more embodiments of the invention are presented below in the description. Other features, objects, and advantages of the invention will be apparent from the description and claims.

DETAILED DESCRIPTION OF THE INVENTION

The prodrugs of S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j ] quinolysin-2-carboxylic acid of Formula I

and their pharmaceutically acceptable solvates, polymorphs or hydrates,

wherein

R is CH ₃ or CH (CH ₃ ) ₂ and

R ₁ is C ₁ -C ₆ alkyl or phenyl, optionally substituted with one or more substituents selected from C ₁ -C ₆ alkyl, halogen, nitro, hydroxy or C ₁ -C ₆ alkoxy.

Solubility in water is an important parameter for the oral bioavailability of a drug. It was found that the water solubility of the compounds of the invention is> 200 mg / ml at pH 7, which is generally greater than that of the free base or hydrochloride.

The compounds of formula I have been shown to provide particularly good absorption reflected in increased AUC and C _max when administered orally in rats compared to the parent compound, i.e., S - (-) - 9-fluoro-6,7-dihydro-8 - (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2-carboxylic acid and its salt with L-arginine. Also, compared with prodrug hydrochloride, S - (-) - 9-fluoro-6,7-dihydro-8- (4-L-alaninyloxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2-carboxylic acid, the compounds of the invention showed increased AUC and C _max when administered orally in rats. In addition, the lower solubility of S - (-) - 9-fluoro-6,7-dihydro-8- (4-L-alaninyloxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo hydrochloride [i, j] -quinolysin-2-carboxylic acid may suffer from a general ionic effect when passing through the gastric region. It has also been found that the compounds of the invention of formula I are well tolerated in large doses in rodents and dogs.

Another embodiment of the present invention relates to a process for preparing compounds of formula I. Compounds of formula I can be prepared by combining an amine protected amino acid with S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1 -yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2-carboxylic acid in the presence of a coupling agent. The amino acid used is L-alanine or L-valine. A compound of the formula is deprotected to obtain a compound of the invention of formula I.

Compounds of the invention can be prepared from S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i , j] quinolysin-2-carboxylic acid by esterification of 4-hydroxypiperidine with N-tert-butoxycarbonyl-L-alanine or N-tert-butoxycarbonyl-L-valine in the presence of a coupling agent using methods known in the art. Typically, the coupling reaction is carried out in the presence of a coupling agent in a suitable solvent and in the presence of one or more bases at a temperature of from -30 ° C to + 150 ° C, to obtain a compound of formula II. Preferred coupling agents include carbodiimides, 2,4,6-trichlorobenzoyl chloride, methanesulfonyl chloride and the like. Carbodiimides, such as dicyclohexylcarbodiimide or N- (3-dimethylaminopropyl) -N-ethylcarbodiimide, can be used as coupling agents. A suitable solvent is selected from halogenated solvents such as dichloromethane, chloroform, or bipolar aprotic solvents such as tetrahydrofuran, N, N-dimethylformamide, or mixtures thereof. One or more bases are selected from triethylamine, N, N-dimethylaminopyridine, Hanig bases (N, N-diisopropylethylamine).

The coupling reaction is carried out by treating S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j ] quinolysin-2-carboxylic acid with N-tert-butoxycarbonyl-L-alanine or N-tert-butoxycarbonyl-L-valine in a halogenated hydrocarbon such as dichloromethane, chloroform or ethylene dichloride and in the presence of N, N-dimethylaminopyridine and a coupling agent, dicyclohexidylcarbodi , at a temperature of from -10 to 0 ° C. Or, alternatively, the coupling reaction can be performed by treating S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2-carboxylic acid N-tert-butoxycarbonyl-L-alanine or N-tert-butoxycarbonyl-L-valine in tetrahydrofuran and dimethylformamide using a combination of trichlorobenzoyl chloride in the presence of N, N- dimethylaminopyridine at a temperature of from -10 to 0 ° C. The coupling reaction may also be carried out by treating S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [ i, j] quinolysin-2-carboxylic acid N-tert-butoxycarbonyl-L-alanine or N-tert-butoxycarbonyl-L-valine in tetrahydrofuran and dimethylformamide using methanesulfonyl chloride as a coupling agent in the presence of triethylamine at a temperature of from -10 to 0 ° C.

The deprotection step of N-tert-butoxycarbonyl (BOC) can be performed using the methods described in TW Greene and PGM Wuts, Protective groups in organic synthesis , 3rd edition, John Wiley & Sons, New York (1999). Accordingly, deprotection can be accomplished by treating a compound of formula II using mildly acidic conditions in aqueous or non-aqueous solutions. Solvents that can be used for the deprotection step in solution are aqueous acid solutions with or without the addition of organic solvents, or non-aqueous organic solvents. Organic solvents are selected from the group consisting of acetone, acetonitrile, ethanol, isopropanol, or mixtures thereof. Acidic conditions can be used using an inorganic acid, such as hydrochloric acid, or, alternatively, using any other organic acid, such as methanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, p- nitrobenzenesulfonic acid, trifluoroacetic acid, acetic acid, formic acid or mixtures thereof. Preferably, the compound of formula II is treated with methanesulfonic acid, p- toluenesulfonic acid, benzenesulfonic acid, p- bromobenzenesulfonic acid or p- nitrobenzenesulfonic acid at a temperature in the range of −10 ° C. to 100 ° C. in acetone or acetonitrile to give a compound of the invention of formula I .

Another aspect of the present invention relates to the purification of the crude product of a compound of the invention of formula I. The purification of the compounds of the invention is carried out by removing impurities, dissolving the impurities in an organic solvent. The solvents used are acetone, diethyl ether or mixtures thereof.

Polymorphs of compounds of formula I can be obtained by crystallization of a compound of formula I under various conditions, for example, temperature, time and / or the use of specific solvents. Hydrates of the compounds of formula I can be prepared by methods known to those skilled in the art.

The term “C ₁ -C ₆ alkyl” refers to straight or branched chain saturated hydrocarbon radicals containing from one to six carbon atoms. Examples of C ₁ -C ₆ alkyl radicals include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl and their branched isomers, such as isopropyl, isobutyl, tert-butyl.

The term “C ₁ -C ₆ alkoxy” refers to an oxygen radical having a substituent in the form of a hydrocarbon chain, wherein the hydrocarbon chain is alkyl (i.e., —O-alkyl). Examples of C ₁ -C ₆ alkoxy are methoxy, ethoxy, propyloxy, isopropyloxy, butyloxy, pentyloxy, hexyloxy.

The term “halogen” as used herein refers to an atom selected from fluoro, chloro, bromo and iodo.

The term “nitro” refers to a group of the formula —NO ₂ -.

The term “alkyl sulfonic acid” refers to the formula (C ₁ -C ₆ alkyl) -SO ₃ H; for example, methanesulfonic acid methane (CH ₃ SO ₃ H), ethanesulfonic acid (CH ₃ CH ₂ SO ₃ H).

The invention also relates to liquid and solid pharmaceutical compositions which contain a prodrug of the invention, such as, for example, injectable solutions, suspensions, emulsions, tablets, coated tablets, coated tablet cores, capsules, solutions, troches, dispersions, pills , granules, suppositories, hard or soft gelatin capsules, and the like.

The pharmaceutical compositions are prepared according to the usual procedures used by those skilled in the art to obtain stable and effective compositions. Such methods include mixing, mixing, suspending, dispersing, emulsifying, dissolving, and the like. active compounds with pharmaceutical auxiliaries or in pharmaceutical auxiliaries, such as a carrier, diluent, solvent or excipient, and processing the components into pharmaceutically suitable forms for parenteral, oral, intranasal, buccal or rectal administration and the like. In solid, liquid, parenteral dosage forms, an effective amount of the active compound or active ingredient is any amount that produces the desired results. In accordance with the present invention, it has been found that the advantageous solubility properties of the prodrugs of the invention can be imparted to the composition of pharmaceutical dosage forms. It can also be used to produce tablets by wet granulation or conventional dry granulation. It can also be used to obtain aqueous dosage forms.

Dosage forms can be obtained by any conventional methods known in the art, but are preferably formulated by mixing the prodrug of the invention with other ingredients. Other ingredients used to formulate solid oral dosage forms include conventional inert ingredients such as microcrystalline cellulose, methyl cellulose and the like, suitable sweeteners, colorants and / or flavors, and, if necessary, their preservatives. Such solid oral dosage forms or dry formulations suitable for preparing suspensions are thus formulated to contain an effective dose of a compound of the invention. In general, solid dosage forms containing from 100 mg to 1500 mg of a compound of the invention are contemplated. Formulations suitable for oral suspension contain a similar dose.

Pharmaceutical formulations can be formulated with adjuvants and additives commonly used in pharmacy, such as tablet binders, excipients, preservatives, tablet disintegrants, flow regulators, emollients, wetting agents, dispersing agents, emulsifiers, solvents, pH modifiers, flavoring agents, and t .P. A second preferred method is parenteral intramuscular, intravenous or subcutaneous administration.

When the pharmaceutical composition is formulated as an injection, all solvents commonly used in the art can be used in formulating the pharmaceutical composition in the form of a solution or suspension. Examples of suitable solvents are water, ethanol, polypropylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters. Sodium chloride, glucose or glycerin may be included in the therapeutic agent. Preferably, the concentration of the active ingredient in the injectable preparation is in the range of 0.1 mg / ml to 100 mg / ml.

In addition to the conventional dosage forms described above, the compounds of the present invention may also be administered by controlled release means and / or delivery devices, such as those described in US Pat. No. 3,845,770; 3,916,899; 3,536,809; 3 598 123 and 4 008 719, the disclosures of which are hereby incorporated by reference.

The full daily dose range is usually from about 200 mg to about 5000 mg of the prodrug of the invention. Preferably, the full daily dose range is from 300 mg to 3000 mg of the prodrug of the invention. However, the dose may be higher or lower depending on the needs and conditions of the patient.

The antibacterial compounds and pharmaceutical compositions of the invention can be used in the treatment of humans and animals having a wide range of bacterial infections, such as impetigo, pneumonia, bronchitis, pharyngitis, endocarditis, urinary tract infections, diabetic foot ulcers, gastrointestinal infections and bacteremia. These bacterial infections can be caused by any of the following bacteria - Staphylococcus aureus , coagulase-negative Staphylococcus aureus, methicillin-resistant Staphylococcus aureus , methicillin-resistant coagulase-negative Staphylococcus aureus, enterococci, beta-hemolytic streptococci, M. streptococci, M. streptococcus infections, M. streptococci multidrug-resistant tuberculosis and other atypical mycobacteria, such as M. intracellulare and M. avium , as well as new gram-negative pathogens, such as Chryseobacterium meningosepticum , Chryseobacteriu m indologense , and other gram-negative pathogens such as E. coli , Klebsiella, Proteus, Serratia, Citrobacter and Pseudomonas.

The present invention also encompasses an anti-infective composition for the treatment of humans and animals for the prevention and / or treatment of systemic infections, especially infections caused by Gram-resistant organisms, infections caused by Gram-negative organisms, mycobacterial infections and infections caused by nosocomial pathogens, which contains a certain amount of a compound of invention, basically sufficient to eliminate the specified infection, but which does not cause any There are undesirable side effects. The compound and compositions of the invention can be administered to humans and animals that are at risk of infection, for example, the compound or composition of the invention can be administered to a patient before and / or after surgery, healthcare professionals or other individuals who are at risk of infection.

The present invention encompasses the administration of the compounds to animals or humans. The compound and compositions used in the invention should accordingly be pharmaceutically acceptable. For the purposes of the invention, such a “pharmaceutically acceptable” component is one which is suitable for use in humans and / or animals without undesirable adverse side effects (such as toxicity, irritation and allergic reaction) commensurate with an acceptable benefit / risk ratio. Animals that can be treated using the compounds of the invention include, but are not limited to, mammals, fish, birds.

The following detailed examples serve to more fully illustrate the invention without limiting its scope. It should be understood that various other embodiments and modifications in the practice of the invention will be obvious to a person skilled in the art and can be easily carried out by him without departing from the scope and spirit of the invention, as described above. Accordingly, the scope of the appended claims is not limited to the precise description given above, but should be construed as including all the features of patentable novelty that are included in the present invention, including all the features and embodiments that a specialist could interpret as equivalents of this invention. The invention is further described in relation to the following experimental work.

The following detailed examples serve to more fully illustrate the invention without limiting its scope.

Experimental part:

(S) -9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] -quinolysin-2-carboxylic acid was obtained according to the procedure described in Chem. Pharm. Bull. 1996, 44 (4), 642-645.

Example 1

Preparation of (2'S, 5S) -9-fluoro-6,7-dihydro-8- (4- (N-tert-butoxycarbonyl-L-alaninyl-hydroxy) -piperidin-1-yl) -5-methyl-1-oxo -1H, 5H-benzo [i, j] quinolysin-2-carboxylic acid

Method 1: To a mixture of N-tert-butoxycarbonyl-L-alanine (473 g) in dichloromethane (2 L), dicyclohexylcarbodiimide (515 g) dissolved in dichloromethane (2 L) was added to give a cloudy suspension. To this cloudy suspension was added 300 g of (S) -9-fluoro-6,7-dihydro-8- (4-hydroxy-piperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i , j] quinolysin-2-carboxylic acid, then 4-N, N-dimethylaminopyridine (58 g), and the reaction mixture was stirred at a temperature of from -10 to 5 ° C for 2 hours. The suspension was filtered and the solid was washed with 500 ml of dichloromethane. The filtrate was washed with water. The filtrate was dried over anhydrous sodium sulfate. The dried organic layer was then concentrated to half its volume, after which a solid precipitated. The solid was filtered off and washed with 300 ml of dichloromethane. The pure organic filtrate was concentrated to dryness to give an oily mass. The oily mass was triturated with diethyl ether (4 L) to give a white solid. The solid was filtered off with suction and washed with diethyl ether (1 L) to give the title compound in an amount of 415 g (94%).

Method 2: To a mixture of triethylamine (98.0 ml) and N-tert-butoxycarbonyl-L-alanine (110 g) in a mixture of tetrahydrofuran (1050 ml) and N, N-dimethylformamide (350 ml) was added 2,4,6- trichlorobenzoyl chloride (100 ml). The resulting mixture was stirred at a temperature of from -5 to 0 ° C for 5 hours. To the reaction mixture were added 4-N, N-dimethylaminopyridine (24 g) and (S) -9-fluoro-6,7-dihydro-8- (4-hydroxy-piperidin-1-yl) -5-methyl-1- oxo-1H, 5H-benzo [i, j] quinolysin-2-carboxylic acid (70 g). The reaction mixture was stirred for another 7 hours at a temperature of from -5 to 0 ° C. The suspension was filtered at room temperature and the filtrate was extracted with ethyl acetate after adding water. Evaporation of the organic layer under reduced pressure gave a sticky solid which, after trituration with diethyl ether, gave a white solid in an amount of 85 g.

Method 3: To a solution of N-tert-butoxycarbonyl-L-alanine (7.9 g) in a mixture of tetrahydrofuran (75 ml) and N, N-dimethylformamide (25 ml) at a temperature of from -10 to 0 ° C was added dropwise methanesulfonyl chloride (2.42 ml). To this solution, triethylamine (8.7 ml) was added dropwise over 5 minutes. The reaction mixture was stirred for 1.5 hours while maintaining the temperature from -10 to 0 ° C. (S) -9-fluoro-6,7-dihydro-8- (4-hydroxy-piperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] was added to the reaction mixture quinolysin-2-carboxylic acid (5.01 g) and 4-N, N-dimethylaminopyridine (1.70 g). The reaction mixture was stirred for an additional 1 hour at a temperature of from -5 to 0 ° C. The suspension was filtered at room temperature and the filtrate was diluted with water (300 ml) and extracted with ethyl acetate (150 ml × 2). Evaporation of the organic layer under reduced pressure gave a sticky solid which, after trituration with diethyl ether, gave a white solid in an amount of 6.38 g (86%).

Example 2

Preparation of (2'S, 5S) -9-fluoro-6,7-dihydro-8- (4-L-alaninyl-hydroxy-piperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo salt [ i, j] quinolysin-2-carboxylic acid with methanesulfonic acid

To a mixture of (2'S, 5S) -9-fluoro-6,7-dihydro-8- (4-N-tert-butoxycarbonyl-L-alaninyloxy-piperidin-1-yl) -5-methyl-1-oxo-1H, 5H-Benzo [i, j] quinolysin-2-carboxylic acid (415 g) in acetone (4.5 L) was added methanesulfonic acid (66 ml). The reaction mixture was stirred at 65-67 ° C overnight. The suspension was filtered at 40-45 ° C. The solid was washed with acetone (1.5 L), then diethyl ether (1.5 L). The off-white solid was dried in vacuum (40-45 mm) at a temperature of 55-60 ° C for a period of 3-4 hours. The target compound was obtained in the form of a free-flowing off-white material 383.0 g (93%).

For MF: C ₂₃ H ₃₀ FN ₃ O ₈ S, MS (ES +) m / z 432 (obtained as free base for MF: C ₂₂ H ₂₆ FN ₃ O ₅ ); MP 278.50 ° C DSC.

Example 3

Preparation of (2'S, 5S) -9-fluoro-6,7-dihydro-8- (4-L-valynyloxy-piperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo salt [i, j] quinolysin-2-carboxylic acid and methanesulfonic acid

Step 1: Preparation of (2'S, 5S) -9-fluoro-6,7-dihydro-8- {4- [N-tert-butyloxycarbonyl-L-valynyloxy] piperidin-1-yl} -5-methyl-1- oxo-1H, 5H-benzo [i, j] quinolysin-2-carboxylic acid

To a mixture of triethylamine (14 ml) and N-tert-butyloxycarbonyl-L-valine (18 g) in a mixture of tetrahydrofuran (50 ml) and N, N-dimethylformamide (15 ml) was added 2,4,6-trichlorobenzoyl chloride (14.3 ml). The resulting mixture was stirred at a temperature of from -5 to 0 ° C for 5 hours. To the reaction mixture was added 4-N, N-dimethylaminopyridine (3.4 g) and (S) -9-fluoro-6,7-dihydro-8- {4-hydroxy-piperidin-1-yl} -5-methyl- 1-oxo-1H, 5H-benzo [i, j] quinolizine-2-carboxylic acid (10 g). The reaction mixture was stirred for another 7 hours at a temperature of from -5 to 0 ° C. The suspension was filtered at room temperature and the filtrate was extracted with ethyl acetate after adding water. Evaporation of the organic layer under reduced pressure gave a sticky solid which, after trituration with diethyl ether, gave a white solid in an amount of 12.5 g.

Step 2: Preparation of (2'S, 5S) -9-fluoro-6,7-dihydro-8- {4-L-valynyloxy-piperidin-1-yl} -5-methyl-1-oxo-1H, 5H-benzo salt [i, j] quinolysin-2-carboxylic acid and methanesulfonic acid

A mixture of the compound obtained in stage 1 (12 g) and methanesulfonic acid (1.81 ml) in acetone (150 ml) was stirred at a temperature of 63-65 ° C for 10 hours. The suspension was filtered at room temperature and the solid was washed with acetone to give an off-white solid in an amount of 9.8 g.

Example 4

Preparation of (2'S, 5S) -9-fluoro-6,7-dihydro-8- {4- (L-alaninyloxy) piperidin-1-yl} -5-methyl-1-oxo-1H, 5H-benzo salt [ i, j] quinolysin-2-carboxylic acid and toluenesulfonic acid

To a mixture of (2'S, 5S) -9-fluoro-6,7-dihydro-8- (4-N-tert-butoxycarbonyl-L-alaninyloxy-piperidin-1-yl) -5-methyl-1-oxo-1H, 5H-Benzo [i, j] quinolysin-2-carboxylic acid (3.0 grams, 5.64 mol) in acetone (35 ml) was added p- toluenesulfonic acid monohydrate (1.61 grams, 8.47 mol). The reaction mixture was refluxed overnight. The suspension was filtered at 40-45 ° C and the resulting solid was washed with acetone, then with diethyl ether. The solid was dried in vacuum (40-45 mm) at temperatures of 55-60 ° C for a period of 3-4 hours, obtaining the target compound in the form of a dirty white solid in the amount of 3.0 grams (yield 88.23%). For MF: C ₂₉ H ₃₄ FN ₃ O ₈ S, MS (M + H) m / z 432 (obtained as free base for MF: C ₂₂ H ₂₆ FN ₃ O ₅ ); MP 232 ° C.

Example 5

Preparation of (2'S, 5S) -9-fluoro-6,7-dihydro-8- {4- (L-alaninyloxy) piperidin-1-yl} -5-methyl-1-oxo-1H, 5H-benzo salt [ i, j] quinolysin-2-carboxylic acid and benzenesulfonic acid

To a mixture of (2'S, 5S) -9-fluoro-6,7-dihydro-8- (4-N-tert-butoxycarbonyl-L-alaninyloxy-piperidin-1-yl) -5-methyl-1-oxo-1H, 5H-Benzo [i, j] quinolysin-2-carboxylic acid (3.0 grams, 5.64 mol) in acetone (35 ml) was added benzenesulfonic acid (1.33 grams, 8.41 mol). The reaction mixture was refluxed overnight. The suspension was filtered at 40-45 ° C and the resulting solid was washed with acetone, then with diethyl ether. The solid was dried in vacuum (40-45 mm) at temperatures of 55-60 ° C for a period of 3-4 hours, obtaining the target compound in the form of a light yellow solid in the amount of 3.1 grams (yield 90.0%). For MF: C ₂₈ H ₃₂ FN ₃ O ₈ S, MS (M + H) m / z 432 (obtained as free base for MF: C ₂₂ H ₂₆ FN ₃ O ₅ ); MP 229 ° C.

Example 6

Preparation of (2'S, 5S) -9-fluoro-6,7-dihydro-8- {4- (L-alaninyloxy) piperidin-1-yl} -5-methyl-1-oxo-1H, 5H-benzo salt [ i, j] quinolysin-2-carboxylic acid and nitrobenzenesulfonic acid

To a mixture of (2'S, 5S) -9-fluoro-6,7-dihydro-8- (4-N-tert-butoxycarbonyl-L-alaninyloxy-piperidin-1-yl) -5-methyl-1-oxo-1H, 5H-Benzo [i, j] quinolysin-2-carboxylic acid (3.0 grams, 5.64 mol) in acetone (35 ml) was added p- nitrobenzenesulfonic acid (1.73 grams, 8.47 mol). The reaction mixture was refluxed overnight. The suspension was filtered at 40-45 ° C and the resulting solid was washed with acetone, then with diethyl ether. The solid was dried in vacuum (40-45 mm) at temperatures of 55-60 ° C for a period of 3-4 hours, obtaining the target compound in the form of a light yellow solid in an amount of 3.3 grams (yield 92.43%). For MF: C ₂₈ H ₃₁ FN ₄ O ₁₀ S, MS (M + H) m / z 432 (obtained as free base for MF: C ₂₂ H ₂₆ FN ₃ O ₅ ); MP 240 ° C.

Test Example 1

Solubility analysis: A method for determining solubility in water at various physiological pH values at temperatures of 25-30 ° C. To a precisely weighed amount of the test substance, a specific buffer solution (parts of approximately 50 microliters each) was added in portions at temperatures of 25-30 ° C. until the solid was completely dissolved and a clear solution was obtained.

Table 1 Solubility at pH 7 Compound Solubility (mg / ml) at
pH 7 (5S) -9-fluoro-6,7-dihydro-8- (4-hydroxy-piperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2- carboxylic acid <1 Salt tetrahydrate (5S) -9-fluoro-6,7-dihydro-8- (4-hydroxy-piperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin- 2-carboxylic acid L-arginine <1 (2'S, 5S) -9-fluoro-6,7-dihydro-8- (4-L-alaninyloxy-piperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo hydrochloride [i, j ] quinolysin-2-carboxylic acid four (2'S, 5S) -9-fluoro-6,7-dihydro-8- (4-L-valynyloxy-piperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2-carboxylic acid <1 Salt (2'S, 5S) -9-fluoro-6,7-dihydro-8- (4-L-alaninyloxy-piperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j ] quinolysin-2-carboxylic acid and methanesulfonic acid > 200 Salt (2'S, 5S) -9-fluoro-6,7-dihydro-8- (4-L-valynyloxy-piperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j ] quinolysin-2-carboxylic acid and methanesulfonic acid > 200

Biological Example

A method for studying pharmacokinetics (PK) in Sprague Dawley rats: a pharmacokinetic study was performed on male Sprague Dawley rats weighing 200-220 g, which were kept on an empty stomach overnight. Salt (2'S, 5S) -9-fluoro-6,7-dihydro-8- (4-L-alaninyloxy-piperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j ] -quinolysin-2-carboxylic acid and methanesulfonic acid (A), (2'S, 5S) -9-fluoro-6,7-dihydro-8- (4-L-alaninyloxy-piperidin-1-yl) -5- hydrochloride methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2-carboxylic acid (B) and the salt (2'S, 5S) -9-fluoro-6,7-dihydro-8- (4-L- valynyloxy-piperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2-carboxylic acid and methanesulfonic acid (C) were dissolved in Milli Q water. Compounds A, B and C was administered at a dose of 50 mg / kg, orally, in terms of the active ingredient. Salt (5S) -9-fluoro-6,7-dihydro-8- (4-hydroxy-piperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolysin-2 -carboxylic acid and L-arginine (D) were dissolved in 1% Tween before use. Compounds A and D were administered at a dose of 200 mg / kg, orally, in terms of the active ingredient. The dose volume was 0.5 ml / 200 g. Blood samples were collected at predetermined time intervals, i.e. 0, 0.5, 1, 2, 4, 6 and 8 hours in eppendorff ampoules containing 10 μl of a saturated solution of sodium fluoride. Plasma was collected after centrifuging a blood sample at 10,000 rpm for 10 minutes. Plasma samples were analyzed by HPLC in order to determine drug levels. The parameters PK AUC, C _{max were} calculated by non-compartmental analysis using Winnonlin software.

table 2 % improvement in C _max and AUC in the study
when administered orally in rats % improvement % increase C _max % increase in AUC A compared to B
50 mg / kg per os 300 175 A compared to D
200 mg / kg per os 200 120 C versus D
50 mg / kg per os 97 92

Claims (18)

1. Compounds having the structure of Formula I

and their pharmaceutically acceptable solvates,
in which R is CH ₃ or CH (CH ₃ ) ₂ ; and
R ₁ is C ₁ -C ₆ alkyl or phenyl, optionally substituted with one or more substituents selected from C ₁ -C ₆ alkyl or nitro. 2. The compounds according to claim 1, in which R ₁ denotes C ₁ -C ₆ alkyl. 3. The compounds of claim 1, wherein R ₁ is phenyl, optionally substituted with one or more substituents selected from C ₁ -C ₆ alkyl or nitro. 4. The compounds according to claim 1, in which R is CH ₃ or CH (CH ₃ ) ₂ and R ₁ is methyl, ethyl, phenyl, p-nitrophenyl, tolyl. 5. A method of obtaining a compound of Formula (I) and its pharmaceutically acceptable solvates, including:
a) a combination of S - (-) - 9-fluoro-6,7-dihydro-8- (4-hydroxypiperidin-1-yl) -5-methyl-1-oxo-1H, 5H-benzo [i, j] quinolizine -2-carboxylic acid with an amine-protected L-alanine or L-valine in one or more solvents to give a protected compound of Formula II;

b) removing the protective group from the compound of Formula II using sulfonic acid of the general Formula R ₁ —SO ₃ H, in which R ₁ has the meaning defined in claim 1;
c) isolation of the compounds of Formula I. 6. The method according to claim 5, in which the coupling reaction is carried out in the presence of a coupling agent. 7. The method according to claim 6, in which the coupling agent includes one or more compounds of dicyclocarbodiimide, 2,4,6-trichlorobenzoyl chloride and methanesulfonyl chloride. 8. The method according to claim 5, further comprising carrying out the coupling reaction in the presence of a base. 9. The method of claim 8, in which the base includes one or more compounds of triethylamine, N, N-dimethylaminopyridine and diisopropylethylamine. 10. The method according to claim 5, in which the solvent includes one or more compounds of dichloromethane, chloroform, ethylene chloride, tetrahydrofuran, N, N-dimethylformamide and mixtures thereof. 11. The method according to claim 5, in which the coupling reaction is carried out at a temperature of from about -10 ° C to about 0 ° C. 12. The method according to claim 5, in which the sulfonic acid used is alkyl sulfonic acid. 13. The method according to item 12, in which the sulfonic acid is methanesulfonic acid. 14. A pharmaceutical composition having antibacterial activity, comprising a therapeutically effective amount of one or more compounds according to claim 1 together with pharmaceutically acceptable carriers, excipients or diluents. 15. The use of compounds according to claim 1 for the manufacture of a medicament for the treatment or prophylaxis of a condition caused or caused by bacterial infections, comprising administering to a mammal a therapeutically effective amount of one or more compounds according to claim 1. 16. The use of compounds according to 15, in which the condition is selected from impetigo, pneumonia, bronchitis, pharyngitis, endocarditis, urinary tract infections, diabetic foot ulcers, gastrointestinal infections or bacteremia. 17. The use of compounds according to clause 15, in which the bacterial infection is caused by gram-positive, gram-negative or anaerobic bacteria. 18. The use according to claim 17, wherein the gram-positive, gram-negative, or anaerobic bacteria are selected from Staphylococcus aureus, coagulase-negative staphylococci, methicillin-resistant Staphylococcus aureus, methicillin-resistant coagulase-negative staphylococci, enterococci viridae streptococci, b. , Mycobacterial infections caused by M. tuberculosis with multidrug resistance and other atypical mycobacteria, such as M. intracellulare and M. avium, as well as new gram-negative pathogens such and as Chryseobacterium meningosepticum, Chryseobacterium indologense, and other gram-negative pathogens such as E. coli, Klebsiella, Proteus, Serratia, Citrobacter and Pseudomonas.